Diphenyl Urea Derivatives

ABSTRACT

The invention relates to novel specifically trifluoromethyl and halogen substituted 1,3-diphenyl ureas and their use as active ingredients in the preparation of pharmaceutical compositions. The invention also concerns related aspects like the preparation of the compounds, pharmaceutical compositions containing one or more of those compounds and especially their use as anti-infectives.

The present invention relates to novel 1,3-diphenyl ureas which arespecifically trifluoromethyl and halogen substituted in the phenylrings, to pharmaceutical compositions containing them and to their usein the treatment and/or prevention of bacterial infections.

In the prior art certain 1,3-diphenyl ureas have been claimed for theirinsecticidal properties (U.S. Pat. No. 2,745,874) or for a generalbiocidal activity for combating insects, fungi and infesting herbs (GBPatent Specification 1 326 481). In one instance, the possibility thatthe insecticidal properties of 1,3-diphenyl ureas might be flanked by abactericidal action, e.g., against S. aureus and fungicidal activity hasbeen described (U.S. Pat. No. 2,745,874). In addition, certain1,3-diphenyl ureas have been described as inhibitors of bacterialRNA-polymerase and this property was claimed to often translate intoantibacterial activity against aerobic Gram-positive and Gram-negativepathogens, e.g., against S. aureus and E. coli (ToIC) (WO 01/51456).

It has now been found that a small group of novel 1,3-diphenyl ureaswith a distinct halogen/trifluoromethyl substitution pattern arespecifically active against bacteria and exhibit virtually no activityagainst fungi and that these novel 1,3-diphenyl ureas are very potentagainst a broad range of aerobic and anaerobic Gram-positive pathogensincluding, among others, multi-drug resistant staphylococci, e.g., S.aureus and S. epidermidis, enterococci, e.g., E. faecalis, streptococci,e.g., S. pneumoniae, S. pyogenes and S. viridans. These propertiesrender these compounds very useful in the treatment of Gram-positivebacterial infections in humans and animals and/or in the decolonizationof sites infested by these pathogens and/or in preventing colonizationof sites from which bacteria can then spread and potentially causebacterial infections. Preferred applications for the compounds of thepresent invention are those related to the topical/localized treatmentof infections in humans and in animals and to the decolonization and/orprevention of colonization of any site which is needed to be renderedsterile from bacteria or in which the bacterial load has to be decreasedto prevent spread of bacteria to other sites and to cause infections.Examples of these applications are treatment of skin, mucosal, ocular,dental, gastro-intestinal and upper respiratory-tract infection,decolonization and/or prevention of bacterial colonization of, amongothers, skin, eyes, nares, mouth, mucosa, gastro-intestinal tract, upperrespiratory tract, prosthetic devices and surfaces in general wherebacteria can survive and eventually replicate e.g., before surgicalpractice and/or in general in any instance in which decolonizationand/or prevention of spread of bacteria to other sites, which bacteriacan infect or colonize, is required.

Therefore, the present invention relates to novel compounds of thegeneral formula I

whereinR¹ and R² represents independently chlorine and bromine;X represents oxygen or sulfur;and pharmaceutically acceptable salts thereof.

Preferred are compounds of formula I wherein R² represents chloro.

Also preferred are compounds where X represents oxygen.

Most preferred compounds of the present invention are:

-   1-(3-Chloro-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,-   1-(3-Bromo-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,    and pharmaceutically acceptable salts thereof.

The expression pharmaceutically acceptable salts encompasses salts witha strong base like an alkali or earth alkali base, e.g. sodiumhydroxide, potassium hydroxide, calcium hydroxide etc., or e.g. cholineetc.

Because of their ability to inhibit aerobic and anaerobic Gram-positivebacteria, compounds of this invention can be used for the treatment ofhuman and animal diseases which are typically associated with one ormore of such type of pathogens and/or in the decolonization of and/or inthe prevention of colonization by one or more of such type of bacteria.This makes compounds of this invention valuable anti-bacterial agents.

The described compounds can be administered by all means known in theart such as, among others, orally, intravenously, topically, rectally,vaginally, sublingually, by inhalation or by any means of local deliverydepending on the site were bacteria are localized as colonizers or asinfecting agents.

Examples of applications are capsules, tablets, orally administeredsuspensions or solutions, suppositories, injections, eye-drops,ointments, aerosols/nebulizers or topical/locally administered forms.Examples of topical forms and of forms suitable for local delivery canbe, among others, gels, creams, ointments, pastes, lotions, solutions,sprays, lozenges, tablets, capsules, sachet, suspension, suppositories,ovules, lacquers, cements, etc. depending on the site that is intendedto treat and/or is intended to reach and/or is intented to protect fromcolonisation, e.g., skin, mucosa, eye, ear, mouth, nares, parts of thegastro-intestinal tract or of the upper-respiratory tract, prostheticdevices.

The described compounds can be also incorporated in the cement and/or inparts of a prosthetic device from which they are released in order toprevent its colonization.

Preferred applications are oraly, topically as well as eye drops.

The dosage used depends upon the type of the specific active ingredient,the use in animal or human, the kind of administration and in case ofapplication in man, the age and the requirements of the patient.Generally, dosages of 0.01-50 mg/kg body weight per day either as asingle or subdivided in 2 to 4 doses per day are considered. For liquidor semi-solid formulations, e.g. solutions, ointments, gels or creams anappropriate amount of a formulation with a ratio between the activeingredient and the excipients in a range between 0.01% to 5% areconsidered. These dosage should be administered preferably in 1 to 4doses per day which are of equal amounts. As usual children shouldreceive lower doses which are adapted to body weight and age.

The preparations with compounds of formula I can contain inertexcipients or also excipients with antibacterial activity. Tablets orgranules, for example, could contain a number of binding agents, fillingexcipients, carrier substances or diluents.

The compositions outlined above may be administered in enteral, oralform or in topical form e.g. as tablets, dragees, gelatine capsules,emulsions, solutions, creams, ointment or suspensions, in intranasalform like sprays or rectally in form of suppositories. These compoundsmay also be administered parenteral, in intramuscular or intraveneousform, e.g. in form of injectable solutions.

These pharmaceutical compositions may contain the compounds of formula Ias well as their pharmaceutically acceptable salts in combination withinorganic and/or organic excipients which are usual in thepharmaceutical industry like lactose, maize or derivatives thereof,talcum, stearinic acid or salts of these materials.

For gelatine capsules vegetable oils, waxes, fats, liquid or half-liquidpolyols etc. may be used. For the preparation of solutions and syrupse.g. water, polyols, saccharose, glucose etc. are used. Injectables areprepared by using e.g. water, polyols, alcohols, glycerin, vegetableoils, lecithin, liposomes etc. Suppositories are prepared by usingnatural or hydrogenated oils, waxes, fatty acids (fats), liquid orhalf-liquid polyols etc. For the preparation of creams, gels, ointmentsetc. for topical and/or local applications e.g. polyols, oils,detergents, penetration enhancer, fillers etc. are used which are knownto someone skilled in the art.

The compositions may contain in addition preservatives, stabilisationimproving substances, viscosity improving or regulating substances,solubility improving substances, sweeteners, dyes, taste improvingcompounds, salts to change the osmotic pressure, buffer, antioxidantsetc.

The compounds of formula I may also be used in co-therapy with one ormore other therapeutics, for example with other classes ofanti-infective agents to increase/complement their anti-infectivespectrum of action, e.g. penicillins and cephalosporins; glycopeptides;quinolones; tetracyclines; aminoglycosides; macrolides, sulfonamidesetc. or antifungals, antiprotozoals etc.

Compounds of this invention can be also incorporated in cleaning and/orcleansing solutions and/or dressings and/or coatings and/or lacquersand/or cements and/or parts of a prosthetic device for decolonizationand/or prevention of bacterial colonization of sites in which bacteriacan survive and eventually replicate causing potential risk forinfections.

Compounds of formula I can be generally synthesized by reacting—asdepicted in Scheme I below—a4-halo-5-(trifluoromethyl)-phenyl-isocyanate or correspondingphenyl-thioisocyanate of formula IIa, with a3-halo-5-trifluoromethyl-aniline IIIa (see also experimental part).Alternatively a 4-halo-5-(trifluoromethyl)-aniline of formula IIb can becoupled with a 3-halo-5-(trifluoromethyl)-phenyl-isocyanate or3-halo-5-(trifluoromethyl)-phenyl-thioisocyanate IIIb to yieldderivatives of the general formula I.

EXAMPLES Abbreviations

DMF: N,N-Dimethyl formamideDMSO: Dimethyl sulfoxideEtOAc: Ethyl acetateMS: Mass spectrometryNMR: Nuclear magnetic resonanceTBME: tert-Butyl methyl ether

THF: Tetrahydrofuran

cHexane: Cyclohexanesat.: saturatedrt: room temperaturer.m.: reaction mixture

Example 1

1-(3-Bromo-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea

To a solution of 3-Bromo-5-trifluoromethyl-aniline (743 μl, 5.25 mmol,1.05 eq.) in dry dichloromethane (20 ml) was added4-Chloro-3-(trifluoromethyl)-phenylisocyanate (1.11 g, 5.0 mmol) at rtand stirred overnight. Then the r.m. was concentrated to circa ⅓ of theinitial volume and the precipitated product was filtered off.Recrystallization from TBME/cHexane yielded 772 mg pure product (33%) asa white powder.

MS (ES⁻): 460.8.

Example 2

1-(3-Chloro-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea

To a solution of 3-Chloro-5-trifluoromethyl-aniline (425 μl, 3.1 mmol,1.03 eq.) in dry THF (15 ml) was added4-Chloro-3-(trifluoromethyl)-phenylisocyanate (665 mg, 3.0 mmol). Afterstirring overnight at rt the mixture was diluted with cHexane (100 ml),washed twice with 2 N HCl, once with sat. NaHCO₃ and brine (100 ml each)and adsorbed on Celite in vacuo. Flash chromatography on silica withcHexane/TBME (4:1 to 2:1) yielded 952 mg product (76%) as a whitepowder.

¹H NMR (DMSO) δ 9.43 (s, 2H), 8.10 (d, J=2.5 Hz, 1H), 7.84 (m, 2H), 7.69(dd, J=9.0, 2.5 Hz, 1H), 7.64 (d, J=9.0 Hz, 1H), 7.45 (s). MS (ES⁻):414.9.

Example 3

1-(3-Bromo-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-thiourea

The title compound was synthesized according to the procedure used inexample 2 starting with 3-Bromo-5-(trifluoromethyl)-aniline and4-Chloro-3-(trifluoromethyl)-phenylisothiocyanate. Yield: 86 mg (36%).

MS (ES⁻): 474.8, 476.8.

Example 4 Biological Results

Antimicrobial susceptibility testing was performed in accordance withthe Clinical and Laboratory Standards Institute (CLSI).

Clinical and Laboratory Standards Institute (CLSI; formerly NCCLS):Methods for Dilution Antimicrobial Susceptibility Tests for BacteriaThat Grow Aerobically; Approved Standard—Seventh Edition (2006).Clinical and Laboratory Standards Institute document M7-A7.

Streptococci (S. pneumoniae, S. pyogenes, S. viridans) were testedfollowing the CLSI methodology with the exception that Todd Hewitt Brothwithout blood was used. Anaerobic bacteria (P. acnes, B. distasonis)were tested following CLSI (formerly NCCLS) guidelines except for usingmicrobroth dilutions in Wilkins Chalgren Broth. National Committee forClinical Laboratory Standards (NCCLS). Methods for Anti-microbialSusceptibility Testing of Anaerobic Bacteria; Approved Standard—SixthEdition (2004). NCCLS document M11-A6.

A) In Vitro Antibacterial Activity of Compounds Against RepresentativePathogens for Nasal Colonization

(Minimum Inhibitory Concentration (MIC) in micrograms/ml) S. aureus S.aureus S. aureus S. epidermidis Example ATCC 25923 MRSA 101 MRSA 39 MRSE70 1 0.5 0.125 0.25 0.25 2 0.25 0.125 0.25 0.25 3 0.125 0.5 0.25 0.25

B) In Vitro Antibacterial Activity of Compounds Against RepresentativePathogens for Ocular Infections

(Minimum Inhibitory Concentration (MIC) in micrograms/ml) S. aureus S.aureus S. aureus S. epidermidis S. pneumoniae S. pyogenes Example 25923MRSA101 MRSA 39 MRSE 70 1/1 GAS-1 1 0.5 0.125 0.25 0.25 0.125 0.25 20.25 0.125 0.25 0.25 0.06 0.25 3 0.125 0.5 0.25 0.25 0.25 0.25

C) In Vitro Antibacterial Activity of Compounds Against RepresentativePathogens for Skin Infections

(Minimum Inhibitory Concentration (MIC) in micrograms/ml) S. E. aureusS. S. faecalis P. MRSA epidermidis pyogenes VanB acnes Example 39 MRSE70 GAS-1 E80-8 6390 1 0.25 0.25 0.25 2 ≦0.03 2 0.25 0.25 0.25 1 ≦0.03 30.25 0.25 0.25 1 ≦0.03

D) In Vitro Antibacterial Activity of Compounds Against RepresentativePathogens for Gastro-Intestinal* or Dental** Infections

(Minimum Inhibitory Concentration (MIC) in micrograms/ml) E. faecalis*B. distasonis* S. viridans** Example VanB E80-8 5770 UHC7 1 2 0.5 0.5 21 1 0.5 3 1 1 1

1. Compounds of the general formula I

wherein R¹ and R² represents independently chlorine and bromine; Xrepresents oxygen or sulfur; and pharmaceutically acceptable saltsthereof.
 2. Compounds of formula I according to claim 1, wherein R²represents chloro.
 3. Compounds according to claim 1, wherein Xrepresents oxygen.
 4. The compounds according to claim 11-(3-Chloro-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,1-(3-Bromo-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,and pharmaceutically acceptable salts thereof.
 5. Pharmaceuticalcompositions for the treatment of infections containing a compound ofclaim 1 and usual carrier materials and adjuvants.
 6. Pharmaceuticalcompositions for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens,containing a compound of claim 1 and usual carrier materials andadjuvants.
 7. The compounds of claim 1 for use as medicaments for thetreatment of infections.
 8. The compounds of claim 1 for use asmedicaments for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens.
 9. Theuse of one or more compounds of claim 1 as active ingredients for theproduction of pharmaceutical compositions for the treatment ofinfections.
 10. The use of one or more compounds of claim 1 as activeingredients for the production of pharmaceutical compositions for thetreatment of infections caused by aerobic and anaerobic Gram positiveand anaerobic Gram negative pathogens.
 11. The use of one or morecompounds of claim 1 as active ingredients for the production ofpharmaceutical compositions for the treatment of bacteria causing nasal,ocular, dental, gastro-intestinal or skin infections.
 12. The use of oneor more compounds of claim 1 as active ingredients for the production ofpharmaceutical compositions for the sterilisation, sanitation,antisepsis, disinfection, decolonisation or prevention of colonisationof the skin, gastro-intestinal tract or the nasal, ocular or dental areaor any type of prosthetic device.
 13. A process for the manufacture ofpharmaceutical compositions for the treatment of infections containingone or more compounds as claimed in claim 1 as active ingredients whichprocess comprises mixing one or more active ingredient withpharmaceutically acceptable excipients in a manner known per se.
 14. Aprocess for the manufacture of pharmaceutical compositions for thetreatment of infections caused by aerobic and anaerobic Gram positiveand anaerobic Gram negative pathogens containing one or more compoundsas claimed in claim 1 as active ingredients which process comprisesmixing one or more active ingredient with pharmaceutically acceptableexcipients in a manner known per se.
 15. Compounds according to claim 2,wherein X represents oxygen.
 16. The compounds according to claim 21-(3-Chloro-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,1-(3-Bromo-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,and pharmaceutically acceptable salts thereof.
 17. The compoundsaccording to claim 31-(3-Chloro-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,1-(3-Bromo-5-trifluoromethyl-phenyl)-3-(4-chloro-3-trifluoromethyl-phenyl)-urea,and pharmaceutically acceptable salts thereof.
 18. Pharmaceuticalcompositions for the treatment of infections containing a compound ofclaim 2 and usual carrier materials and adjuvants.
 19. Pharmaceuticalcompositions for the treatment of infections containing a compound ofclaim 3 and usual carrier materials and adjuvants.
 20. Pharmaceuticalcompositions for the treatment of infections containing a compound ofclaim 4 and usual carrier materials and adjuvants.
 21. Pharmaceuticalcompositions for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens,containing a compound of claim 2 and usual carrier materials andadjuvants.
 22. Pharmaceutical compositions for the treatment ofinfections caused by aerobic and anaerobic Gram positive and anaerobicGram negative pathogens, containing a compound of claim 3 and usualcarrier materials and adjuvants.
 23. Pharmaceutical compositions for thetreatment of infections caused by aerobic and anaerobic Gram positiveand anaerobic Gram negative pathogens, containing a compound of claim 4and usual carrier materials and adjuvants.
 24. The compounds of claim 2for use as medicaments for the treatment of infections.
 25. Thecompounds of claim 3 for use as medicaments for the treatment ofinfections.
 26. The compounds of claim 4 for use as medicaments for thetreatment of infections.
 27. The compounds of claim 2 for use asmedicaments for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens.
 28. Thecompounds of claim 3 for use as medicaments for the treatment ofinfections caused by aerobic and anaerobic Gram positive and anaerobicGrain negative pathogens.
 29. The compounds of claim 4 for use asmedicaments for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens.
 30. Theuse of one or more compounds of claim 2 as active ingredients for theproduction of pharmaceutical compositions for the treatment ofinfections.
 31. The use of one or more compounds of claim 3 as activeingredients for the production of pharmaceutical compositions for thetreatment of infections.
 32. The use of one or more compounds of claim 4as active ingredients for the production of pharmaceutical compositionsfor the treatment of infections.
 33. The use of one or more compounds ofclaim 2 as active ingredients for the production of pharmaceuticalcompositions for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens.
 34. Theuse of one or more compounds of claim 3 as active ingredients for theproduction of pharmaceutical compositions for the treatment ofinfections caused by aerobic and anaerobic Gram positive and anaerobicGram negative pathogens.
 35. The use of one or more compounds of claim 4as active ingredients for the production of pharmaceutical compositionsfor the treatment of infections caused by aerobic and anaerobic Grampositive and anaerobic Gram negative pathogens.
 36. The use of one ormore compounds of claim 2 as active ingredients for the production ofpharmaceutical compositions for the treatment of bacteria causing nasal,ocular, dental, gastro-intestinal or skin infections.
 37. The use of oneor more compounds of claim 3 as active ingredients for the production ofpharmaceutical compositions for the treatment of bacteria causing nasal,ocular, dental, gastro-intestinal or skin infections.
 38. The use of oneor more compounds of claim 4 as active ingredients for the production ofpharmaceutical compositions for the treatment of bacteria causing nasal,ocular, dental, gastro-intestinal or skin infections.
 39. The use of oneor more compounds of claim 2 as active ingredients for the production ofpharmaceutical compositions for the sterilisation, sanitation,antisepsis, disinfection, decolonisation or prevention of colonisationof the skin, gastro-intestinal tract or the nasal, ocular or dental areaor any type of prosthetic device.
 40. The use of one or more compoundsof claim 3 as active ingredients for the production of pharmaceuticalcompositions for the sterilisation, sanitation, antisepsis,disinfection, decolonisation or prevention of colonisation of the skin,gastro-intestinal tract or the nasal, ocular or dental area or any typeof prosthetic device.
 41. The use of one or more compounds of claim 4 asactive ingredients for the production of pharmaceutical compositions forthe sterilisation, sanitation, antisepsis, disinfection, decolonisationor prevention of colonisation of the skin, gastro-intestinal tract orthe nasal, ocular or dental area or any type of prosthetic device.
 42. Aprocess for the manufacture of pharmaceutical compositions for thetreatment of infections containing one or more compounds as claimed inclaim 2 as active ingredients which process comprises mixing one or moreactive ingredient with pharmaceutically acceptable excipients in amanner known per se.
 43. A process for the manufacture of pharmaceuticalcompositions for the treatment of infections containing one or morecompounds as claimed in claim 3 as active ingredients which processcomprises mixing one or more active ingredient with pharmaceuticallyacceptable excipients in a manner known per se.
 44. A process for themanufacture of pharmaceutical compositions for the treatment ofinfections containing one or more compounds as claimed in claim 4 asactive ingredients which process comprises mixing one or more activeingredient with pharmaceutically acceptable excipients in a manner knownper se.
 45. A process for the manufacture of pharmaceutical compositionsfor the treatment of infections caused by aerobic and anaerobic Grampositive and anaerobic Gram negative pathogens containing one or morecompounds as claimed in claim 2 as active ingredients which processcomprises mixing one or more active ingredient with pharmaceuticallyacceptable excipients in a manner known per se.
 46. A process for themanufacture of pharmaceutical compositions for the treatment ofinfections caused by aerobic and anaerobic Gram positive and anaerobicGram negative pathogens containing one or more compounds as claimed inclaim 3 as active ingredients which process comprises mixing one or moreactive ingredient with pharmaceutically acceptable excipients in amanner known per se.
 47. A process for the manufacture of pharmaceuticalcompositions for the treatment of infections caused by aerobic andanaerobic Gram positive and anaerobic Gram negative pathogens containingone or more compounds as claimed in claim 4 as active ingredients whichprocess comprises mixing one or more active ingredient withpharmaceutically acceptable excipients in a manner known per se.